DESCRIPTION (from applicant's abstract): Otitis media is the most common diagnosis in pediatric patients who visit physicians for illness in the United States. Despite the prevalence of otitis media, its potential for morbidity, the enormous health care expenditures resulting from its treatment and the increasing therapeutic challenges imposed by antimicrobial resistance, much is still unknown about the cellular and molecular immunologic and inflammatory events in this disease process. Middle car epithelium is known to play a significant role in the pathophysiology of otitis media through a cytokine mediated inflammatory response. Recent advances in cell culture techniques have enabled investigations of middle ear epithelium in culture and provide a mechanism to further our understanding of the cellular and molecular events in otitis media. The goals of this project are to test the hypothesis that cultured middle ear epithelial cells (MEEC) exposed to the inflammatory cytokines tumor necrosis factor-alpha. (TNF-alpha), interleukin- 1 beta (IL- 1 beta), interleukin-6 (IL- 6) and interleukin-8 (IL-8) will exhibit changes in: 1) mucoglycoprotein secretion (MGP), 2) mucin gene expression and 3) morphology. Using cell and molecular biology techniques, differences in epithelial response to this array of inflammatory cytokines will be compared and the ability to block the specific changes brought about by these cytokines by the use of cytokine inhibitors will be examined. Post-receptor signal transduction of TNF-alpha, IL- 1 beta, IL-6 and IL-8 in regards to MEEC MGP secretion will also be examined. Understanding the cellular and molecular events involved in cytokine-middle ear epithelial interactions, MGP production, mucin gene expression and signaling pathways will potentially allow for novel and efficacious treatments for otitis media through modulation of the cytokine pathway and MGP production by middle ear epithelial cells.